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food security aspects related to the environmental release of pharmaceuticals
Chemosphere xxx (2014) xxx–xxx
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Food safety/food security aspects related to the environmental release of pharmaceuticals Gianfranco Brambilla a,⇑, Cecilia Testa b a b
Istituto Superiore di Sanità, Toxicological Chemistry Unit, Rome, Italy Istituto Zooprofilattico Sperimentale della Sardegna, Lab Drug Residues, Sassari, Italy
h i g h l i g h t s
g r a p h i c a l a b s t r a c t
Top soil and surface water intake
expose grazing animals to contaminants. Pharmaceuticals in the environment may be a risk for food safety/security. Proposed EQS in surface waters also seem appropriate for food safety assessment. The use of biosolids on pasture may represent a risk in free-grazing species. The definition of EQS in soil to support food safety and food security is envisaged.
a r t i c l e
i n f o
Article history: Received 11 December 2013 Received in revised form 22 January 2014 Accepted 22 January 2014 Available online xxxx Keywords: Pharmaceuticals Top soil Surface waters Animal production Food safety Food security
a b s t r a c t The environmental presence of pharmaceuticals in top soil and in water where extensive animal farming occurs may represent an involuntary source of residues in food that might affect both food safety and food security. We modelled the presence of residues in animal matrices from the inventoried environmental concentration of selected drugs in surface waters (range: 0.1–10 lg L 1) and agriculture soils (range: 1–100 lg kg 1 dry weight), accounting for animal production parameters (i.e., forages, water intake and milk and egg production) and drug pharmacokinetics. The results indicate that the contamination of tetracyclines in top soil may represent a major issue both for the compliance with maximum residue levels in food (100–300 ng g 1) and for the claim of organic products. via surface water, animals may be vulnerable to the intake of anabolics and growth-promoting agents, such as 17-beta estradiol and clenbuterol, only under a worst-case scenario. Their identification, which is currently achievable at a pg g 1 level in animal specimens, is considered proof of illegal treatment and can lead to the prosecution of farmers. The Environmental Quality Standards that have been proposed for priority substances in surface waters may also be considered protective in terms of food security/food safety; however, a broadspectrum characterisation of drugs within the agriculture context could be envisaged to refine the uncertainties in the risk assessment and for combined intakes. Ó 2014 Elsevier Ltd. All rights reserved.
⇑ Corresponding author. Address: Istituto Superiore di Sanità, Toxicological Chemistry Unit, Viale Regina Elena, 299, I-00161 Rome, Italy. Tel.: +39 6 4990 2764; fax: +39 6 4990 2836. E-mail addresses: [email protected] (G. Brambilla), cecilia.testa@ izs-sardegna.it (C. Testa).
1. Introduction There is consolidated evidence that intensive farming systems (fattening calves, pigs, poultry, and aquaculture) represent a source
http://dx.doi.org/10.1016/j.chemosphere.2014.01.024 0045-6535/Ó 2014 Elsevier Ltd. All rights reserved.
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G. Brambilla, C. Testa / Chemosphere xxx (2014) xxx–xxx
Nomenclature BATs Best Available Techniques bw body weight COR carry-over rate d day dm dry matter E2 17 beta estradiol EE2 ethinyl estradiol EFSA European Food Safety Authority EMA (EMEA) European Medicine Agency
for the release of pharmaceuticals in the environment (Jjemba, 2002). Thus far, drugs with antimicrobial/anti-parasitic activity are the most practicable tools to limit the onset and spread of infectious diseases to high-density feedlots (EU Commission, 2010). Antimicrobials in medicated feeds and, to a lesser extent, in feed additive at doses up to g kg 1 in feed, as in the case of sulphonamides and tetracyclines, determine the presence of drug residues at the mg kg 1 level in animal dejects. Manure and slurry are then dressed on agricultural soils as fertilisers (Migliore et al., 2003, 2010); thus, determining the environmental release is also extended to endogenous hormones from pig breeding farms (Pinheiro et al., 2013). In the case of off-shore aquaculture farms, veterinary drugs and their metabolites are released directly into water bodies (Cabello, 2006). Therefore, registered veterinary drugs and additives should comply with an Environmental Risk Assessment during the authorisation procedure, and possible adverse effects on biota should be reported under pharmacovigilance programmes underpinned in the EU Commission Directive 2001/ 82/EC. Moreover, intensive animal farming settlements could apply to a voluntary Eco-Management and Audit Scheme (EMAS) (EU Commission Decision, 2011), i.e., through the application of the Best Available Techniques (BATs) (EU Commission, 2013), to mitigate the environmental impact of their activities. In recent years, the evidence regarding the impact of the environmental quality on food-producing animals and their products in rural and extensive farming systems has progressively arisen. Priority contaminants, such as heavy metals and persistent organic pollutants, may affect the compliance with the food safety legislation of products from grazing cattle and pigs (Edwards, 2003; Gummow et al., 2006; Smith et al., 2009), free range flocks, (Van Overmeire et al., 2009), and extensively farmed fish (Miniero et al., 2013). In such cases, the animals’ intake of contaminants that are associated with natural resources (top soil and the related fauna), grass, surface water and associated sediments, and air deposition on fodders may result in greater relevance than that associated with commercial feeds. In this paper, we attempt to assess possible environmental quality parameters for the presence of pharmaceuticals in the agriculture zones that are devoted to extensive farming systems, both in terms of food safety (levels of authorised drugs in veterinary medicine below the maximum residue limit in edible tissues) and food security (presence of unauthorised/undeclared drugs below their analytical thresholds), as far as the involuntary presence of food residues of an undeclared/unauthorised drug may severely affect the socio-economics of the farming system, according to the food safety legislation in place within the European Union. Within the sustainability policies to reduce the carbon- and water-footprints in food production systems, the proposed use of bio-solids as top soil enhancers in agriculture (Chen et al., 2012) and the watering of fodders from reclaimed waters (Arnold et al., 2012), which both originate from urban waste water treatments plants, as well as the direct animal intake of pharmaceuticals from fresh-
EMAS Eco Management and Audit Scheme EQS Environmental Quality Standard EU European Union LC–MS/MS liquid chromatography–tandem mass spectrometry LoD limit of determination MoSS margin of food safety/security MRL maximum residue limit WWTP waste water treatment plant
water, may represent an emerging source of xenobiotics into the food chain. 2. Materials and methods 2.1. Pharmaceuticals in top soil and surface water are of relevance for food safety/food security The group of tetracyclines (oxy-, chlor-, tetra-, doxycycline) have been considered because these drugs are among the most prescribed antimicrobials in both veterinary and human medicine and because of their ability to bind humic acids and metals that are present in top soil (Hamscher et al., 2002; Blackwell et al., 2009). Their concentration ranges in agricultural soils have been derived from the international literature, which was reviewed by the Bio Intelligence Service (2013). Because of their wide consumption in both human and veterinary medicine and because of their relevance in pharmacoresistance outbreaks, along with their ability to bind to soil, fluoroquinolones were also accounted for (Leal et al., 2013). For surface waters, the group of sulphonamides, as drugs that are more prone to be released in WWTP effluents, have been chosen according to Loos et al., 2008. Moreover, the concentration of anti-asthmatic drugs, such as albuterol/salbutamol and clenbuterol, have also been considered due to their proposition in the recent past as growth promoting agents in feedlots; their use is currently forbidden in the European Union and is severely prosecuted and fined (EU Directive 96/23/EC, Annex I). Under this perspective, estrogens, such as ethinyl estradiol (EES) and 17 beta estradiol (E2), which are either listed in the above quoted Annex A or next to be framed within the European Parliament Directive 2013/39/EC as priority substances for their potential harmful effects in the aquatic environment, have also been accounted for. Our attention has also been focused on diclofenac, which is a non-steroidal anti-inflammatory drug licensed in veterinary medicine (EMA, 2009) but of potential eco-toxicological concern in water bodies (Johnson et al., 2013). Other pharmaceuticals, such as thiabendazole, which is an anti-parasitic and anti-fungal drug that is commonly used in animals and in agriculture (Correa and Escandar, 2006), and lorazepam, which is a short-acting benzodiazepine from human prescriptions (Mendoza et al., 2013), have been included in this assessment because of the reported concentrations above 1 lg L 1 in surface waters, which may represent a potential risk in animal food production systems. The amounts, which refer to the sales, of the main classes of antimicrobials that are prescribed in veterinary medicine and that are considered environmental contaminants in top soil and surface water are reported in Fig. 1. 2.2. Top soil intake in grazing animals Several papers examine top soil intake in farmed animals, with reported differences among species and seasonal variations
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G. Brambilla, C. Testa / Chemosphere xxx (2014) xxx–xxx
fluoroquinolones
sulphonamides
tetracyclines
0
500
1000
1500
2000
2500
3000
3500
tonnes of active ingredients Fig. 1. Sales, in tonnes of active ingredients, of veterinary antimicrobials agents, in EU Countries, in 2011. Data from the Third ESVAC report (EMA/236501/2013).
accounting for the latitude of the pasture. In rural flocks, the top soil intake (worms included) could represent up to 50% of the feed ratio, on a dry matter basis (Waegeneers et al., 2009). Grazing sheep could assume, with grass, up to 30% of top soil (Smith et al., 2009); in a recent opinion on ‘‘dioxin’’ and ‘‘dioxin-like’’ substances in sheep liver, the European Food Safety Authority (EFSA, 2011) considered 6% as the median of the median values reported in the literature. Similarly, top soil intakes in cows and pigs may represent up to 20% of the overall dry matter daily intake (Edwards, 2003; Gummow et al., 2006). In our model, a conservative approach has been followed, considering a soil intake at 50% and 20% of the dry matter intake from the feed regimen of poultry (hens and broilers) and sheep, respectively. 2.3. Fodder uptake of pharmaceuticals from agriculture soils The uptake of pharmaceuticals from soils to grass and fodders that are intended for animal nutrition has not been considered quantitatively when referring to the epigeal parts of the plant (Boxall et al., 2006; Migliore et al., 2010; Kang et al., 2013; Chitescu et al., 2013) and has been computed at 5% of the concentration found in top soil, on a dry matter basis. Therefore, the contribution to the overall environmental intake of selected pharmaceuticals from grass was estimated from the previously inventoried top soil contamination and corrected for the above-mentioned transfer factor, which accounts for the daily dry matter intake of grass. 2.4. Animal production and pharmacokinetic parameters Default animal production parameters for broilers, laying hens, growing pigs, lactating sheep and cows, which have been derived from an animal production manual that referred to Mediterranean areas (Bittante et al., 2005), are reported in Table 1. Regarding such
production parameters, drug carry-over rates (COR) have been computed as the ratio between the total amount of the drug excreted or present in the target tissue and the total amount of the drug involuntarily ingested via top soil or water. The COR of sulphonamides to milk (contaminated at 20 ng mL 1 for an estimated 40 L d 1 production) have been derived from Agarwal (1992), who reported sub-therapeutic exposures (140 mg head 1 d 1) in dairy cows because of the drugs carry-over from medicated to non-medicated feeds that were processed on the same line in feed mills. Similarly, the carry-over of tetracyclines to the eggs and liver of laying hens and broilers has been derived from Vandenberge et al. (2012a,b), under the assumption of a daily intake of 100 g (broilers) and 120 g (hens) contaminated feed, accounting for a daily 50 g egg production and a liver weight of 36 g in a 3.0 kg bw broiler. Enrofloxacin and its metabolite, ciprofloxacin, were assumed to be representative of the fluoroquinolones antimicrobial category; their pharmacokinetic parameters in the milk and liver of dairy animals and in the eggs and liver of poultry were derived from EMEA (1998), Cornejo et al. (2011), and Kalpana et al. (2012). The pharmacokinetic parameters for thiabendazole and for diclofenac were derived from the corresponding European Medicine Agency fact sheets (EMA, 2004, 2009), whereas those pharmacokinetic parameters referring to illicit drugs (estrogenic compounds, beta adrenergic agonists) in animal production were derived from Gleixner (1998) and Pleadin et al. (2011), respectively. Because of the lack of data regarding the administration of benzodiazepines in dairy animals, the relevant parameters have been derived from human pharmacology (Lemmer et al., 2007).
3. Results The outcomes of the modelled environmental quality parameters for the intake of selected pharmaceuticals from surface waters and top soil as environmental food are illustrated in Table 2, along with the pharmacokinetic parameters that were considered. The legislative maximum residue levels in food that are currently in place within the European Union are listed in Table 3, along with the limit of determinations, accounting for the currently achievable limits of identification. Fig. 2 illustrates the margin of safety/margin of security (MoSS), which is computed as the ratio between the modelled concentrations in the selected animal matrices (Table 2) and the correspondent analytical threshold (MRL/LoD) that is reported in Table 3. Values <1 indicate a potential risk for food security and/or food safety. The indirect contribution to sheep milk contamination from the uptake of grass contaminated with oxytetracycline (top soil: 10– 100 mg kg 1) resulted in the range of 0.4–4.0 ng g 1 as a result of a grass intake as dry matter of 2800 g per head d 1, a COR grass/ soil = 0.05 on a dry basis, and a COR grass-to-milk of 0.0057. For the considered fluoroquinolone concentration in top soil, such contribution was assumed negligible.
Table 1 Animal production parameters accounting for the carry-over rates from surface waters and top soil, in selected animal production. 1
Species
Body weight (kg)
Water intake + 23 °C (L head d
)
Top soil intake (g head d
Dairy cow Dairy sheep
550 60
102 10
2300 840
Laying hen Broilers Fattening pig
3.5 3.0 80
0.15 0.20 5
60 50 nc
Veal calves Veal calves
80 80
5 5
nc nc
1
)
Product (g)
Matrix
Notes
23 000 2000 1200 50 60 1600 2 3000 2800
Milk Milk Liver Egg Liver Liver Choroid Urine Serum
Daily average production Daily average production 2% of the bw Daily average production 2% of the bw 2% of the bw Pigmented layer 75% of the water intake 3.5% of the bw
bw = body weight; nc = not considered.
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G. Brambilla, C. Testa / Chemosphere xxx (2014) xxx–xxx
Table 2 Residue levels modelled in food producing animal specimens, because of the environmental intake of selected pharmaceuticals via surface water or via amended top soil. Range (lg g
**
1
Intake (lg)
COR
Min
Max
100
60–6000
0.0033
4
400
1
100
50–5000
0.0576
48
4800
Top soil
1
100
840–840 000
0.012
8
840
Tetracyclines
Top soil
1
100
840–840 000
0.0057
2
240
Fluoroquinolones
Top soil
0.01
1
0.6–60
0.002
0.024
2.4
Fluoroquinolones
Top soil
0.01
1
0.5–50
0.002
0.016
1.6
Fluoroquinolones
Top soil
0.01
1
8.4–840
0.001
0.004
0.4
Intake (lg)
COR
Residue range (ng g
Source
Min
Max
Tetracyclines
Top soil
1
Tetracyclines
Top soil
Tetracyclines
Range (lg L
*
1
Drug
dry weight)
1
)
Min
Max
Residue range (ng g
1
)
)
Min
Max
Sulphonamides
Water
0.1
10
10.2–1020
0.0057
0.002
0.253
Etinyl estradiol
Water
0.035*
3.5*
0.35–35**
0.9
<.001
0.010
Beta estradiol
Water
0.04*
4.0*
0.8–80**
0.9
<0.001
0.027
Albuterol
Water
0.01
1
0.05–5
0.006
<0.001
0.015
Clenbuterol
Water
0.001
0.1
0.005–0.5
0.8
0.001
0.133
Diclofenac
Water
0.1
10
10.2–1020
0.023
0.010
1.02
Lorazepam
Water
0.05
0.150
5 100–15 300
0.007
0.001
0.003
Thiabendazole
Water
0.05
5
5.1–5100
0.0033
<0.001
0.073
Values expressed as ng L Values expressed as ng.
Matrix/animal
Eggs Laying hens Liver Broilers Liver Sheep Milk Sheep Eggs Laying hens Liver Broilers Liver Sheep Matrix/animal
Milk Cows Urine Veal calves Serum Veal calves Choroid Pig Urine Veal calves Milk Cows Milk Cows Milk Cows
1
.
Table 3 Maximum Residue Limits (MRL) and LOD of selected pharmaceuticals in the considered matrix. Substance
Matrix
MRL*
LoD*
Tetracycline
Milk Egg Liver
100 200 300
1 1 1
Oxytetracycline
Milk Egg Liver
100 200 300
1 1 1
Chlortetracycline
Milk Egg Liver
100 200 300
1 1 1
Doxycycline
Milk Egg Liver
100 200 300
1 1 1
Sulphonamides Salbutamol Ethinyl estradiol
Milk Coroid Urine
100 10 na
0.03 0.25 0.01
Reference
LC MS/MS Apley (2011)
na = not applicable. Values expressed as lg kg
*
LC MS/MS Arroyo-Manzanares et al. (2014) LC MS/MS Agilent (2011) LC MS/MS Zhao et al. (2013)
1
.
4. Discussion 4.1. Uncertainties The following uncertainties in the model can be recognised for an appropriate discussion. No data are available regarding the unintentional presence of pharmaceuticals in the surface water
that is administered to feedlots. In some rural districts, such as those districts that are close to river plains, the animal water supply is obtained directly from lakes and rivers, possibly under the pressure of effluents from civil wastewater treatment plants. Therefore, under a conservative scenario, it seemed worth considering a drug data set that refers to highly impacted surface water (Blair et al., 2013; Oldenkamp et al., 2013). No direct measure-
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G. Brambilla, C. Testa / Chemosphere xxx (2014) xxx–xxx
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for a grass production of 7.5 ton year 1 as dry matter). The large variability inventoried in the top soil intake from grazing animals (EFSA, 2011) also accounts for seasonal and geographic differences. In Southern Europe during the summer season, a higher top soil intake may be expected due to reduced grass coverage of the soil. In contrast, the water intake in dairy animals usually shows seasonal variation, with a 20% increase during the summer season, which is characterised by heat waves that may reach air temperatures up to 37 °C. Simultaneously, seasonal variations may affect drug concentrations in water, which accounts for water in surface water basins and for the seasonal prescribing of drugs, such as antibiotics during the winter and anti-asthmatic during the spring seasons. In this case, the uncertainties may affect estimates in a neutral way, although a quite large dispersion around the modelled residues may be expected, following seasonal trends. The drug bioavailabilities from soil and their related pharmacokinetics have been assumed equivalent to those values described in poultry and dairy animals that have been exposed to cross-contaminated feeds on the basis that their intake is almost contemporary to that from grass, worms, and seeds. For diclofenac, thus far, no pharmacokinetic parameters are available for oral consumption in dairy cows; therefore, we used the parameters that referred to the administration by the intramuscular route (EMA, 2009), which corrected for 50% of the bioavailability, with expert judgement. The pharmacokinetic model of fluoroquinolones accounts for the cumulative excretion of the parent drug enrofloxacin and its main metabolite, ciprofloxacin. Due to the poor stability of tetracyclines in water, the model we propose assumed that their intake was negligible from such a source; similarly, due to the leakage of sulphonamides from soil, the intake of sulpha drugs from top soil was estimated as irrelevant (Blackwell et al., 2009). For plants grown on amended soils, the intake of the epigeal part has been considered largely predominant with respect to that of roots (usually 10-fold more contaminated than the epigeal parts) (Migliore et al., 2010). 4.2. Pharmaceutical intake from surface waters
Fig. 2. Margin of safety/security (MoSS) from the top for tetracyclines, sulphonamides, and salbutamol, in selected matrices from food producing animals, accounting for best/worst case exposure scenarios (grey and dark bars), as reported in Table 2. For sulphonamides, a central scenario was considered, as a matter of possible cumulative exposure.
ments are available for top soil contamination with pharmaceuticals at the grazing time either. Because it is a common practice to graze herds and flocks 35–60 d after the pasture dressing with biosolids, and accounting for the stability of oxytetracycline in amended soils (Wang and Yates, 2008), the concentrations reported in the literature have been assumed as a realistic scenario. However, a progressive decrease in the exposure can be considered when sheep are allowed to graze three months after the soil amendment due to the degradation of the drugs, particularly in the presence of rainfall, which is able to improve soil moisture above 20%. Another mitigation factor in the intake of tetracyclines, and in general for those drugs that are associated with the top soil, may rely on the flock/herd density, which inversely correlates with food residue levels (Waegeneers et al., 2009). Therefore, the uncertainty should be addressed towards a lower animal exposure. However, a factor that accounts for the biological mass dilution of the contamination has been not considered for sheep, under the assumption of an irrelevant impact on extensive farming systems (150–200 kg body mass ha 1, equivalent to 3–4 sheep ha 1
From the modelled exposure that is reported in Table 3 for sulphonamides, even under a MRL of 100 ng g 1 set on cumulative basis in milk, which accounts for all of the contributions from human and veterinary prescriptions belonging to this therapeutic class, the reported concentrations in water would not represent a risk of incompliance according to the food safety legislation. The Environmental Quality Standards (EQS) of 0.4 ng L 1 proposed for E2 in water by the European Commission also seems adequately protective in terms of food safety/security; E2 serum levels above 0.04 ng mL 1 are considered proof of illegal anabolic treatments in veal calves, as far as such a threshold is significantly higher than native levels that are present in pre-pubertal cattle (Brambilla et al., 2003). A possible concern may derive from the 4 ng L 1 concentration because of those concentrations that have been reported in run-off water close to intensive breeding farms (Pinheiro et al., 2013); the estimated serum concentration of 0.03 ng mL 1 is rather close to the above reported threshold value. EE2 is a synthetic oestrogen; thus, its presence in animal matrices has no threshold level. Its identification at the lowest level reachable, which accounts for the present analytical performance, is considered proof of illegal treatment (EU Regulation 882/2004). According to the currently achievable limits of identification, levels in urine at a few pg mL 1 following the water intake at the EQS concentration of 0.035 ng L 1 can hardly be detected under routine residue analysis (Table 2). Therefore, the proposed EQS must also be considered safe for food production. Anti-asthmatic drugs have been extensively used as leaning agents in meat production in cattle and pigs, with acute intoxication outbreaks in humans following the ingestion of contaminated
Please cite this article in press as: Brambilla, G., Testa, C. Food safety/food security aspects related to the environmental release of pharmaceuticals. Chemosphere (2014), http://dx.doi.org/10.1016/j.chemosphere.2014.01.024
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meat and liver (Wu et al., 2013). Therefore, their use has been progressively forbidden in feedlots; their presence in pigmented matrices, such as choroid, due to the strong binding with melanin is also considered a proof of illegal treatment, although the choroid cannot be ranked among edible tissues (Sauer et al., 1995). The reported concentration of albuterol/salbutamol in surface water may represent a potential risk for unintentional exposure in feedlots, as far as the estimated residual levels are in line with the analytical performances in use in veterinary forensic medicine (Table 3, Fig. 2). For diclofenac, with a proposed EQS in surface waters of 0.1 lg L 1, the residual level at the MRL can be achieved only in highly impacted anthropogenic areas, as far as its prescription in veterinary medicine rarely encompasses a large animal number due to its administration via the intra-muscular route. Additionally, in this case, the proposed EQS can be considered aligned to the food safety legislation, with respect to MRLs, apart from potential allergy outbreaks induced in sensitised consumers after the intake of food residues at a pg g 1 level in milk. Thiabendazole is a worldwide known drug belonging to the class of benzimidazoles, which is used in both veterinary medicine as an anti-parasitic drug and in agriculture as an anti-fungal agent. Thiabendazole has been reported to reach up to 1.44 mg L 1 in the Parana river (Correa and Escandar, 2006). Accounting for an estimated carry-over rate from water to milk of 0.0033, such a concentration, which is well above the EQS of 50 lg L 1, would not pose any risk for food safety in dairy products, accounting for a MRL of 100 ng mL 1 in milk. 4.3. Pharmaceuticals in top soil The direct intake of oxytetracycline from top soil in all grazing animals that were considered may represent both a matter for food safety (exceeding MRLs) and food security (presence of undeclared antimicrobials in organic products), as far as the estimated contamination in milk, liver, and eggs fall well above the analytical performances of the methods. The indirect intake from contaminated grass seems irrelevant as far as this contamination could contribute up to 5.7% of the overall contamination estimated in milk. The outcomes for fluoroquinolones are different from those outcomes modelled for tetracyclines. The reported environmental concentrations in soil (0.02–2 mg kg 1 dm) are irrelevant for determining potential food safety noncompliance with respect to MRLs. However, the environmental presence of drugs belonging to such a class in the environment should be adequately evaluated within pharmacovigilance programmes that are focused on anti-microbial resistance outbreaks. 5. Conclusions From the modelled impact of the environmental quality of surface waters on food safety, the proposed EQS for the considered veterinary drugs in surface waters does not appear to represent a matter in terms of the violation of the corresponding maximum residue limits in food. However, a broad-spectrum characterisation via multi-residue methods in water for all of the drugs belonging to the same pharmacological class, which, thus, embraces all possible contributions from veterinary and human prescriptions, should be envisaged to reduce the uncertainties. Possible noncompliance that affects food security may arise from the environmental presence of unauthorised drugs, where the threshold levels for the compliance are represented by the currently achievable limits of identification, in the range of pg g 1. In contrast, the impact of the drugs that are potentially present in top soil enhancers may also be recognised as
the main risk factor as a consequence of the increasing efficacy in the WWPT treatments to remove xenobiotics from water effluents. A proper characterisation of organic contamination in bio-solids and in composts intended for agriculture use, which is also because of an improved abatement of the pharmaceutical load, should be acknowledged as a priority for food safety and food security, within an overall guaranteed environmental quality that is able to support rural and organic animal farming. This support may improve the impact of the rural and organic farming on socio-economics, because the added economic value of the organic food retains in the local community, but also for the beneficial effects on other activities, such as tourism, to create greater economic opportunities in a wider community. Acknowledgements The Grant from the Italian Ministry of Health, RF 2009 No. 1534860, Project ENVI-FOOD, is acknowledged. References Agarwal, V.K. (Ed.), 1992. Analysis of antibiotic/drug residues in food products of animal origin. In: Proc. American Chemical Society Agricultural and Food Chemistry Division Symposium on Antibiotic-drug residues in food products of animal origin, 26–30 August, New York. Plenum Press, New York, 263 pp. (1992 VII Edition). Agilent application note 5990-8788EN-2011 Determination of beta2 agonists in pork with SPE clean-up and LC–MS/MS detection. Available at:. Apley, M. NPB #09-257 Chlortetracycline, Oxytetracycline, Tetracycline and Bacitracin Tissue Residue Studies in Swine Conducted in Reference to Foreign Export Markets –, , PharmCATS Bioanalytical Services, Department of Clinical Sciences College of Veterinary Medicine, Kansas State University, 2011, available at: . Arnold, R.G., Saez, A.E., Snyder, S., Maeng, S.K., Lee, C., Woods, G.J., Li, X., Choi, H., 2012. Direct potable reuse of reclaimed wastewater: it is time for a rational discussion. Rev. Environ. Health 27, 197–206. Arroyo-Manzanares, N., Gámiz-Gracia, L., García-Campaña, A.M., 2014. Alternative sample treatments for the determination of sulphonamides in milk by HPLC with fluorescence detection. Food Chem. 143, 459–464. Bio Intelligence Service, 2013. Study on the risk of environmental effects of medicinal products. Draft Final Report prepared for Executive Agency for Health and Consumers. Bittante, G., Andrighetto, I., Ramanzin, M., 2005. Tecniche di produzione animale (in italian). Liviana editrice, Padua (Italy), ISBN: 978884947083. Blackwell, P.A., Kay, P., Ashauer, R., Boxall, A.B., 2009. Effects of agricultural conditions on the leaching behaviour of veterinary antibiotics in soils. Chemosphere 75 (1), 13–19. http://dx.doi.org/10.1016/ j.chemosphere.2008.11.070, Epub 2009 January 6. Blair, B.D., Crago, J.P., Hedman, C.J., Klaper, R.D., 2013. Pharmaceuticals and personal care products found in the Great Lakes above concentrations of environmental concern. Chemosphere 93, 2116–2123. Boxall, A.B.A., Johnson, P., Smith, E.J., Sinclair, C.J., Stutt, E., Levy, L., 2006. Uptake of veterinary medicines from soils into plants. J. Agric. Food Chem. 54, 2288–2297. Brambilla, G., Ballerini, A., Civitareale, C., Fiori, M., Neri, B., Cavallina, R., Nardoni, A., Giannetti, L., 2003. Oxidative stress as a bio-marker of estrogen exposure in healthy veal calves. Anal. Chim. Acta 483, 281–288. Cabello, F.C., 2006. Heavy use of prophylactic antibiotics in aquaculture: a growing problem for human and animal health and for the environment. Environ. Microbiol. 8, 1137–1144. Chen, H., Yan, S.H., Ye, Z.L., Meng, H.J., Zhu, Y.G., 2012. Utilization of urban sewage sludge: Chinese perspectives. Environ. Sci. Pollut. Res. Int. 19, 1454–1463. Chitescu, C.L., Nicolau, A.I., Stolker, A.A.M., 2013. Uptake of oxytetracycline, sulfamethoxazole and ketoconazole from fertilised soils by plants. Food Add. Contamin.: Part A 30 (6), 1138–1146. http://dx.doi.org/10.1080/ 19440049.2012.725479. Cornejo, J., Lapierre, L., Iragüen, D., Cornejo, S., Cassus, G., Richter, P., San Martín, B., 2011. Study of enrofloxacin and flumequine residues depletion in eggs of laying hens after oral administration. J. Vet. Pharmacol. Therap. 35, 67–72. Correa, R.A., Escandar, G.M., 2006. A new analytical application of nylon-induced room-temperature phosphorescence: determination of thiabendazole in water samples. Anal. Chim. Acta 571, 58–65. Edwards, S.A., 2003. Intake of nutrients from pasture by pigs (Review). Proc. Nutr. Soc. 62, 257–265. European Food Safety Authority (EFSA) Panel on Contaminants in the Food Chain, 2011. Scientific Opinion on the risk to public health related to the presence of high levels of dioxins and dioxin-like PCBs in liver from sheep and deer EFSA Journal 9, 2297 [71 pp.]. http://dx.doi.org/10.2903/j.efsa.2011.2297 Available online:
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Contents lists available at ScienceDirect
Chemosphere journal homepage: www.elsevier.com/locate/chemosphere
Food safety/food security aspects related to the environmental release of pharmaceuticals Gianfranco Brambilla a,⇑, Cecilia Testa b a b
Istituto Superiore di Sanità, Toxicological Chemistry Unit, Rome, Italy Istituto Zooprofilattico Sperimentale della Sardegna, Lab Drug Residues, Sassari, Italy
h i g h l i g h t s
g r a p h i c a l a b s t r a c t
Top soil and surface water intake
expose grazing animals to contaminants. Pharmaceuticals in the environment may be a risk for food safety/security. Proposed EQS in surface waters also seem appropriate for food safety assessment. The use of biosolids on pasture may represent a risk in free-grazing species. The definition of EQS in soil to support food safety and food security is envisaged.
a r t i c l e
i n f o
Article history: Received 11 December 2013 Received in revised form 22 January 2014 Accepted 22 January 2014 Available online xxxx Keywords: Pharmaceuticals Top soil Surface waters Animal production Food safety Food security
a b s t r a c t The environmental presence of pharmaceuticals in top soil and in water where extensive animal farming occurs may represent an involuntary source of residues in food that might affect both food safety and food security. We modelled the presence of residues in animal matrices from the inventoried environmental concentration of selected drugs in surface waters (range: 0.1–10 lg L 1) and agriculture soils (range: 1–100 lg kg 1 dry weight), accounting for animal production parameters (i.e., forages, water intake and milk and egg production) and drug pharmacokinetics. The results indicate that the contamination of tetracyclines in top soil may represent a major issue both for the compliance with maximum residue levels in food (100–300 ng g 1) and for the claim of organic products. via surface water, animals may be vulnerable to the intake of anabolics and growth-promoting agents, such as 17-beta estradiol and clenbuterol, only under a worst-case scenario. Their identification, which is currently achievable at a pg g 1 level in animal specimens, is considered proof of illegal treatment and can lead to the prosecution of farmers. The Environmental Quality Standards that have been proposed for priority substances in surface waters may also be considered protective in terms of food security/food safety; however, a broadspectrum characterisation of drugs within the agriculture context could be envisaged to refine the uncertainties in the risk assessment and for combined intakes. Ó 2014 Elsevier Ltd. All rights reserved.
⇑ Corresponding author. Address: Istituto Superiore di Sanità, Toxicological Chemistry Unit, Viale Regina Elena, 299, I-00161 Rome, Italy. Tel.: +39 6 4990 2764; fax: +39 6 4990 2836. E-mail addresses: [email protected] (G. Brambilla), cecilia.testa@ izs-sardegna.it (C. Testa).
1. Introduction There is consolidated evidence that intensive farming systems (fattening calves, pigs, poultry, and aquaculture) represent a source
http://dx.doi.org/10.1016/j.chemosphere.2014.01.024 0045-6535/Ó 2014 Elsevier Ltd. All rights reserved.
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Nomenclature BATs Best Available Techniques bw body weight COR carry-over rate d day dm dry matter E2 17 beta estradiol EE2 ethinyl estradiol EFSA European Food Safety Authority EMA (EMEA) European Medicine Agency
for the release of pharmaceuticals in the environment (Jjemba, 2002). Thus far, drugs with antimicrobial/anti-parasitic activity are the most practicable tools to limit the onset and spread of infectious diseases to high-density feedlots (EU Commission, 2010). Antimicrobials in medicated feeds and, to a lesser extent, in feed additive at doses up to g kg 1 in feed, as in the case of sulphonamides and tetracyclines, determine the presence of drug residues at the mg kg 1 level in animal dejects. Manure and slurry are then dressed on agricultural soils as fertilisers (Migliore et al., 2003, 2010); thus, determining the environmental release is also extended to endogenous hormones from pig breeding farms (Pinheiro et al., 2013). In the case of off-shore aquaculture farms, veterinary drugs and their metabolites are released directly into water bodies (Cabello, 2006). Therefore, registered veterinary drugs and additives should comply with an Environmental Risk Assessment during the authorisation procedure, and possible adverse effects on biota should be reported under pharmacovigilance programmes underpinned in the EU Commission Directive 2001/ 82/EC. Moreover, intensive animal farming settlements could apply to a voluntary Eco-Management and Audit Scheme (EMAS) (EU Commission Decision, 2011), i.e., through the application of the Best Available Techniques (BATs) (EU Commission, 2013), to mitigate the environmental impact of their activities. In recent years, the evidence regarding the impact of the environmental quality on food-producing animals and their products in rural and extensive farming systems has progressively arisen. Priority contaminants, such as heavy metals and persistent organic pollutants, may affect the compliance with the food safety legislation of products from grazing cattle and pigs (Edwards, 2003; Gummow et al., 2006; Smith et al., 2009), free range flocks, (Van Overmeire et al., 2009), and extensively farmed fish (Miniero et al., 2013). In such cases, the animals’ intake of contaminants that are associated with natural resources (top soil and the related fauna), grass, surface water and associated sediments, and air deposition on fodders may result in greater relevance than that associated with commercial feeds. In this paper, we attempt to assess possible environmental quality parameters for the presence of pharmaceuticals in the agriculture zones that are devoted to extensive farming systems, both in terms of food safety (levels of authorised drugs in veterinary medicine below the maximum residue limit in edible tissues) and food security (presence of unauthorised/undeclared drugs below their analytical thresholds), as far as the involuntary presence of food residues of an undeclared/unauthorised drug may severely affect the socio-economics of the farming system, according to the food safety legislation in place within the European Union. Within the sustainability policies to reduce the carbon- and water-footprints in food production systems, the proposed use of bio-solids as top soil enhancers in agriculture (Chen et al., 2012) and the watering of fodders from reclaimed waters (Arnold et al., 2012), which both originate from urban waste water treatments plants, as well as the direct animal intake of pharmaceuticals from fresh-
EMAS Eco Management and Audit Scheme EQS Environmental Quality Standard EU European Union LC–MS/MS liquid chromatography–tandem mass spectrometry LoD limit of determination MoSS margin of food safety/security MRL maximum residue limit WWTP waste water treatment plant
water, may represent an emerging source of xenobiotics into the food chain. 2. Materials and methods 2.1. Pharmaceuticals in top soil and surface water are of relevance for food safety/food security The group of tetracyclines (oxy-, chlor-, tetra-, doxycycline) have been considered because these drugs are among the most prescribed antimicrobials in both veterinary and human medicine and because of their ability to bind humic acids and metals that are present in top soil (Hamscher et al., 2002; Blackwell et al., 2009). Their concentration ranges in agricultural soils have been derived from the international literature, which was reviewed by the Bio Intelligence Service (2013). Because of their wide consumption in both human and veterinary medicine and because of their relevance in pharmacoresistance outbreaks, along with their ability to bind to soil, fluoroquinolones were also accounted for (Leal et al., 2013). For surface waters, the group of sulphonamides, as drugs that are more prone to be released in WWTP effluents, have been chosen according to Loos et al., 2008. Moreover, the concentration of anti-asthmatic drugs, such as albuterol/salbutamol and clenbuterol, have also been considered due to their proposition in the recent past as growth promoting agents in feedlots; their use is currently forbidden in the European Union and is severely prosecuted and fined (EU Directive 96/23/EC, Annex I). Under this perspective, estrogens, such as ethinyl estradiol (EES) and 17 beta estradiol (E2), which are either listed in the above quoted Annex A or next to be framed within the European Parliament Directive 2013/39/EC as priority substances for their potential harmful effects in the aquatic environment, have also been accounted for. Our attention has also been focused on diclofenac, which is a non-steroidal anti-inflammatory drug licensed in veterinary medicine (EMA, 2009) but of potential eco-toxicological concern in water bodies (Johnson et al., 2013). Other pharmaceuticals, such as thiabendazole, which is an anti-parasitic and anti-fungal drug that is commonly used in animals and in agriculture (Correa and Escandar, 2006), and lorazepam, which is a short-acting benzodiazepine from human prescriptions (Mendoza et al., 2013), have been included in this assessment because of the reported concentrations above 1 lg L 1 in surface waters, which may represent a potential risk in animal food production systems. The amounts, which refer to the sales, of the main classes of antimicrobials that are prescribed in veterinary medicine and that are considered environmental contaminants in top soil and surface water are reported in Fig. 1. 2.2. Top soil intake in grazing animals Several papers examine top soil intake in farmed animals, with reported differences among species and seasonal variations
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fluoroquinolones
sulphonamides
tetracyclines
0
500
1000
1500
2000
2500
3000
3500
tonnes of active ingredients Fig. 1. Sales, in tonnes of active ingredients, of veterinary antimicrobials agents, in EU Countries, in 2011. Data from the Third ESVAC report (EMA/236501/2013).
accounting for the latitude of the pasture. In rural flocks, the top soil intake (worms included) could represent up to 50% of the feed ratio, on a dry matter basis (Waegeneers et al., 2009). Grazing sheep could assume, with grass, up to 30% of top soil (Smith et al., 2009); in a recent opinion on ‘‘dioxin’’ and ‘‘dioxin-like’’ substances in sheep liver, the European Food Safety Authority (EFSA, 2011) considered 6% as the median of the median values reported in the literature. Similarly, top soil intakes in cows and pigs may represent up to 20% of the overall dry matter daily intake (Edwards, 2003; Gummow et al., 2006). In our model, a conservative approach has been followed, considering a soil intake at 50% and 20% of the dry matter intake from the feed regimen of poultry (hens and broilers) and sheep, respectively. 2.3. Fodder uptake of pharmaceuticals from agriculture soils The uptake of pharmaceuticals from soils to grass and fodders that are intended for animal nutrition has not been considered quantitatively when referring to the epigeal parts of the plant (Boxall et al., 2006; Migliore et al., 2010; Kang et al., 2013; Chitescu et al., 2013) and has been computed at 5% of the concentration found in top soil, on a dry matter basis. Therefore, the contribution to the overall environmental intake of selected pharmaceuticals from grass was estimated from the previously inventoried top soil contamination and corrected for the above-mentioned transfer factor, which accounts for the daily dry matter intake of grass. 2.4. Animal production and pharmacokinetic parameters Default animal production parameters for broilers, laying hens, growing pigs, lactating sheep and cows, which have been derived from an animal production manual that referred to Mediterranean areas (Bittante et al., 2005), are reported in Table 1. Regarding such
production parameters, drug carry-over rates (COR) have been computed as the ratio between the total amount of the drug excreted or present in the target tissue and the total amount of the drug involuntarily ingested via top soil or water. The COR of sulphonamides to milk (contaminated at 20 ng mL 1 for an estimated 40 L d 1 production) have been derived from Agarwal (1992), who reported sub-therapeutic exposures (140 mg head 1 d 1) in dairy cows because of the drugs carry-over from medicated to non-medicated feeds that were processed on the same line in feed mills. Similarly, the carry-over of tetracyclines to the eggs and liver of laying hens and broilers has been derived from Vandenberge et al. (2012a,b), under the assumption of a daily intake of 100 g (broilers) and 120 g (hens) contaminated feed, accounting for a daily 50 g egg production and a liver weight of 36 g in a 3.0 kg bw broiler. Enrofloxacin and its metabolite, ciprofloxacin, were assumed to be representative of the fluoroquinolones antimicrobial category; their pharmacokinetic parameters in the milk and liver of dairy animals and in the eggs and liver of poultry were derived from EMEA (1998), Cornejo et al. (2011), and Kalpana et al. (2012). The pharmacokinetic parameters for thiabendazole and for diclofenac were derived from the corresponding European Medicine Agency fact sheets (EMA, 2004, 2009), whereas those pharmacokinetic parameters referring to illicit drugs (estrogenic compounds, beta adrenergic agonists) in animal production were derived from Gleixner (1998) and Pleadin et al. (2011), respectively. Because of the lack of data regarding the administration of benzodiazepines in dairy animals, the relevant parameters have been derived from human pharmacology (Lemmer et al., 2007).
3. Results The outcomes of the modelled environmental quality parameters for the intake of selected pharmaceuticals from surface waters and top soil as environmental food are illustrated in Table 2, along with the pharmacokinetic parameters that were considered. The legislative maximum residue levels in food that are currently in place within the European Union are listed in Table 3, along with the limit of determinations, accounting for the currently achievable limits of identification. Fig. 2 illustrates the margin of safety/margin of security (MoSS), which is computed as the ratio between the modelled concentrations in the selected animal matrices (Table 2) and the correspondent analytical threshold (MRL/LoD) that is reported in Table 3. Values <1 indicate a potential risk for food security and/or food safety. The indirect contribution to sheep milk contamination from the uptake of grass contaminated with oxytetracycline (top soil: 10– 100 mg kg 1) resulted in the range of 0.4–4.0 ng g 1 as a result of a grass intake as dry matter of 2800 g per head d 1, a COR grass/ soil = 0.05 on a dry basis, and a COR grass-to-milk of 0.0057. For the considered fluoroquinolone concentration in top soil, such contribution was assumed negligible.
Table 1 Animal production parameters accounting for the carry-over rates from surface waters and top soil, in selected animal production. 1
Species
Body weight (kg)
Water intake + 23 °C (L head d
)
Top soil intake (g head d
Dairy cow Dairy sheep
550 60
102 10
2300 840
Laying hen Broilers Fattening pig
3.5 3.0 80
0.15 0.20 5
60 50 nc
Veal calves Veal calves
80 80
5 5
nc nc
1
)
Product (g)
Matrix
Notes
23 000 2000 1200 50 60 1600 2 3000 2800
Milk Milk Liver Egg Liver Liver Choroid Urine Serum
Daily average production Daily average production 2% of the bw Daily average production 2% of the bw 2% of the bw Pigmented layer 75% of the water intake 3.5% of the bw
bw = body weight; nc = not considered.
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G. Brambilla, C. Testa / Chemosphere xxx (2014) xxx–xxx
Table 2 Residue levels modelled in food producing animal specimens, because of the environmental intake of selected pharmaceuticals via surface water or via amended top soil. Range (lg g
**
1
Intake (lg)
COR
Min
Max
100
60–6000
0.0033
4
400
1
100
50–5000
0.0576
48
4800
Top soil
1
100
840–840 000
0.012
8
840
Tetracyclines
Top soil
1
100
840–840 000
0.0057
2
240
Fluoroquinolones
Top soil
0.01
1
0.6–60
0.002
0.024
2.4
Fluoroquinolones
Top soil
0.01
1
0.5–50
0.002
0.016
1.6
Fluoroquinolones
Top soil
0.01
1
8.4–840
0.001
0.004
0.4
Intake (lg)
COR
Residue range (ng g
Source
Min
Max
Tetracyclines
Top soil
1
Tetracyclines
Top soil
Tetracyclines
Range (lg L
*
1
Drug
dry weight)
1
)
Min
Max
Residue range (ng g
1
)
)
Min
Max
Sulphonamides
Water
0.1
10
10.2–1020
0.0057
0.002
0.253
Etinyl estradiol
Water
0.035*
3.5*
0.35–35**
0.9
<.001
0.010
Beta estradiol
Water
0.04*
4.0*
0.8–80**
0.9
<0.001
0.027
Albuterol
Water
0.01
1
0.05–5
0.006
<0.001
0.015
Clenbuterol
Water
0.001
0.1
0.005–0.5
0.8
0.001
0.133
Diclofenac
Water
0.1
10
10.2–1020
0.023
0.010
1.02
Lorazepam
Water
0.05
0.150
5 100–15 300
0.007
0.001
0.003
Thiabendazole
Water
0.05
5
5.1–5100
0.0033
<0.001
0.073
Values expressed as ng L Values expressed as ng.
Matrix/animal
Eggs Laying hens Liver Broilers Liver Sheep Milk Sheep Eggs Laying hens Liver Broilers Liver Sheep Matrix/animal
Milk Cows Urine Veal calves Serum Veal calves Choroid Pig Urine Veal calves Milk Cows Milk Cows Milk Cows
1
.
Table 3 Maximum Residue Limits (MRL) and LOD of selected pharmaceuticals in the considered matrix. Substance
Matrix
MRL*
LoD*
Tetracycline
Milk Egg Liver
100 200 300
1 1 1
Oxytetracycline
Milk Egg Liver
100 200 300
1 1 1
Chlortetracycline
Milk Egg Liver
100 200 300
1 1 1
Doxycycline
Milk Egg Liver
100 200 300
1 1 1
Sulphonamides Salbutamol Ethinyl estradiol
Milk Coroid Urine
100 10 na
0.03 0.25 0.01
Reference
LC MS/MS Apley (2011)
na = not applicable. Values expressed as lg kg
*
LC MS/MS Arroyo-Manzanares et al. (2014) LC MS/MS Agilent (2011) LC MS/MS Zhao et al. (2013)
1
.
4. Discussion 4.1. Uncertainties The following uncertainties in the model can be recognised for an appropriate discussion. No data are available regarding the unintentional presence of pharmaceuticals in the surface water
that is administered to feedlots. In some rural districts, such as those districts that are close to river plains, the animal water supply is obtained directly from lakes and rivers, possibly under the pressure of effluents from civil wastewater treatment plants. Therefore, under a conservative scenario, it seemed worth considering a drug data set that refers to highly impacted surface water (Blair et al., 2013; Oldenkamp et al., 2013). No direct measure-
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G. Brambilla, C. Testa / Chemosphere xxx (2014) xxx–xxx
5
for a grass production of 7.5 ton year 1 as dry matter). The large variability inventoried in the top soil intake from grazing animals (EFSA, 2011) also accounts for seasonal and geographic differences. In Southern Europe during the summer season, a higher top soil intake may be expected due to reduced grass coverage of the soil. In contrast, the water intake in dairy animals usually shows seasonal variation, with a 20% increase during the summer season, which is characterised by heat waves that may reach air temperatures up to 37 °C. Simultaneously, seasonal variations may affect drug concentrations in water, which accounts for water in surface water basins and for the seasonal prescribing of drugs, such as antibiotics during the winter and anti-asthmatic during the spring seasons. In this case, the uncertainties may affect estimates in a neutral way, although a quite large dispersion around the modelled residues may be expected, following seasonal trends. The drug bioavailabilities from soil and their related pharmacokinetics have been assumed equivalent to those values described in poultry and dairy animals that have been exposed to cross-contaminated feeds on the basis that their intake is almost contemporary to that from grass, worms, and seeds. For diclofenac, thus far, no pharmacokinetic parameters are available for oral consumption in dairy cows; therefore, we used the parameters that referred to the administration by the intramuscular route (EMA, 2009), which corrected for 50% of the bioavailability, with expert judgement. The pharmacokinetic model of fluoroquinolones accounts for the cumulative excretion of the parent drug enrofloxacin and its main metabolite, ciprofloxacin. Due to the poor stability of tetracyclines in water, the model we propose assumed that their intake was negligible from such a source; similarly, due to the leakage of sulphonamides from soil, the intake of sulpha drugs from top soil was estimated as irrelevant (Blackwell et al., 2009). For plants grown on amended soils, the intake of the epigeal part has been considered largely predominant with respect to that of roots (usually 10-fold more contaminated than the epigeal parts) (Migliore et al., 2010). 4.2. Pharmaceutical intake from surface waters
Fig. 2. Margin of safety/security (MoSS) from the top for tetracyclines, sulphonamides, and salbutamol, in selected matrices from food producing animals, accounting for best/worst case exposure scenarios (grey and dark bars), as reported in Table 2. For sulphonamides, a central scenario was considered, as a matter of possible cumulative exposure.
ments are available for top soil contamination with pharmaceuticals at the grazing time either. Because it is a common practice to graze herds and flocks 35–60 d after the pasture dressing with biosolids, and accounting for the stability of oxytetracycline in amended soils (Wang and Yates, 2008), the concentrations reported in the literature have been assumed as a realistic scenario. However, a progressive decrease in the exposure can be considered when sheep are allowed to graze three months after the soil amendment due to the degradation of the drugs, particularly in the presence of rainfall, which is able to improve soil moisture above 20%. Another mitigation factor in the intake of tetracyclines, and in general for those drugs that are associated with the top soil, may rely on the flock/herd density, which inversely correlates with food residue levels (Waegeneers et al., 2009). Therefore, the uncertainty should be addressed towards a lower animal exposure. However, a factor that accounts for the biological mass dilution of the contamination has been not considered for sheep, under the assumption of an irrelevant impact on extensive farming systems (150–200 kg body mass ha 1, equivalent to 3–4 sheep ha 1
From the modelled exposure that is reported in Table 3 for sulphonamides, even under a MRL of 100 ng g 1 set on cumulative basis in milk, which accounts for all of the contributions from human and veterinary prescriptions belonging to this therapeutic class, the reported concentrations in water would not represent a risk of incompliance according to the food safety legislation. The Environmental Quality Standards (EQS) of 0.4 ng L 1 proposed for E2 in water by the European Commission also seems adequately protective in terms of food safety/security; E2 serum levels above 0.04 ng mL 1 are considered proof of illegal anabolic treatments in veal calves, as far as such a threshold is significantly higher than native levels that are present in pre-pubertal cattle (Brambilla et al., 2003). A possible concern may derive from the 4 ng L 1 concentration because of those concentrations that have been reported in run-off water close to intensive breeding farms (Pinheiro et al., 2013); the estimated serum concentration of 0.03 ng mL 1 is rather close to the above reported threshold value. EE2 is a synthetic oestrogen; thus, its presence in animal matrices has no threshold level. Its identification at the lowest level reachable, which accounts for the present analytical performance, is considered proof of illegal treatment (EU Regulation 882/2004). According to the currently achievable limits of identification, levels in urine at a few pg mL 1 following the water intake at the EQS concentration of 0.035 ng L 1 can hardly be detected under routine residue analysis (Table 2). Therefore, the proposed EQS must also be considered safe for food production. Anti-asthmatic drugs have been extensively used as leaning agents in meat production in cattle and pigs, with acute intoxication outbreaks in humans following the ingestion of contaminated
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G. Brambilla, C. Testa / Chemosphere xxx (2014) xxx–xxx
meat and liver (Wu et al., 2013). Therefore, their use has been progressively forbidden in feedlots; their presence in pigmented matrices, such as choroid, due to the strong binding with melanin is also considered a proof of illegal treatment, although the choroid cannot be ranked among edible tissues (Sauer et al., 1995). The reported concentration of albuterol/salbutamol in surface water may represent a potential risk for unintentional exposure in feedlots, as far as the estimated residual levels are in line with the analytical performances in use in veterinary forensic medicine (Table 3, Fig. 2). For diclofenac, with a proposed EQS in surface waters of 0.1 lg L 1, the residual level at the MRL can be achieved only in highly impacted anthropogenic areas, as far as its prescription in veterinary medicine rarely encompasses a large animal number due to its administration via the intra-muscular route. Additionally, in this case, the proposed EQS can be considered aligned to the food safety legislation, with respect to MRLs, apart from potential allergy outbreaks induced in sensitised consumers after the intake of food residues at a pg g 1 level in milk. Thiabendazole is a worldwide known drug belonging to the class of benzimidazoles, which is used in both veterinary medicine as an anti-parasitic drug and in agriculture as an anti-fungal agent. Thiabendazole has been reported to reach up to 1.44 mg L 1 in the Parana river (Correa and Escandar, 2006). Accounting for an estimated carry-over rate from water to milk of 0.0033, such a concentration, which is well above the EQS of 50 lg L 1, would not pose any risk for food safety in dairy products, accounting for a MRL of 100 ng mL 1 in milk. 4.3. Pharmaceuticals in top soil The direct intake of oxytetracycline from top soil in all grazing animals that were considered may represent both a matter for food safety (exceeding MRLs) and food security (presence of undeclared antimicrobials in organic products), as far as the estimated contamination in milk, liver, and eggs fall well above the analytical performances of the methods. The indirect intake from contaminated grass seems irrelevant as far as this contamination could contribute up to 5.7% of the overall contamination estimated in milk. The outcomes for fluoroquinolones are different from those outcomes modelled for tetracyclines. The reported environmental concentrations in soil (0.02–2 mg kg 1 dm) are irrelevant for determining potential food safety noncompliance with respect to MRLs. However, the environmental presence of drugs belonging to such a class in the environment should be adequately evaluated within pharmacovigilance programmes that are focused on anti-microbial resistance outbreaks. 5. Conclusions From the modelled impact of the environmental quality of surface waters on food safety, the proposed EQS for the considered veterinary drugs in surface waters does not appear to represent a matter in terms of the violation of the corresponding maximum residue limits in food. However, a broad-spectrum characterisation via multi-residue methods in water for all of the drugs belonging to the same pharmacological class, which, thus, embraces all possible contributions from veterinary and human prescriptions, should be envisaged to reduce the uncertainties. Possible noncompliance that affects food security may arise from the environmental presence of unauthorised drugs, where the threshold levels for the compliance are represented by the currently achievable limits of identification, in the range of pg g 1. In contrast, the impact of the drugs that are potentially present in top soil enhancers may also be recognised as
the main risk factor as a consequence of the increasing efficacy in the WWPT treatments to remove xenobiotics from water effluents. A proper characterisation of organic contamination in bio-solids and in composts intended for agriculture use, which is also because of an improved abatement of the pharmaceutical load, should be acknowledged as a priority for food safety and food security, within an overall guaranteed environmental quality that is able to support rural and organic animal farming. This support may improve the impact of the rural and organic farming on socio-economics, because the added economic value of the organic food retains in the local community, but also for the beneficial effects on other activities, such as tourism, to create greater economic opportunities in a wider community. Acknowledgements The Grant from the Italian Ministry of Health, RF 2009 No. 1534860, Project ENVI-FOOD, is acknowledged. References Agarwal, V.K. (Ed.), 1992. Analysis of antibiotic/drug residues in food products of animal origin. In: Proc. American Chemical Society Agricultural and Food Chemistry Division Symposium on Antibiotic-drug residues in food products of animal origin, 26–30 August, New York. Plenum Press, New York, 263 pp. (1992 VII Edition). Agilent application note 5990-8788EN-2011 Determination of beta2 agonists in pork with SPE clean-up and LC–MS/MS detection. Available at:
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G. Brambilla, C. Testa / Chemosphere xxx (2014) xxx–xxx European Medicines Agency (EMA), 1998. Committee for medicinal products for veterinary use: Enrofloxacin. Summary report EMEA/MRL/389/98-FINAL. European Medicines Agency (EMA), 2004. Committee for medicinal products for veterinary use: thiabendazole. Summary report EMEA/MRL/868/03-FINAL. European Medicines Agency (EMA), 2009. Committee for medicinal products for veterinary use: European publicMRL assessment report (EPMAR) Diclofenac: Bovine milk Doc. Ref: EMEA/CVMP/67421/2009. European Commission, 2010. Evaluation of the EU Legislative Framework in the Field of Medicated Feed. Final Report available at:
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